Halo Tag: Difference between revisions
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the dye back to its non-fluorescent state. | the dye back to its non-fluorescent state. | ||
[[Image:Principle of HaloTag.jpg | | [[Image:Principle of HaloTag.jpg | frame | upright= 1.5 |none | alt= | Fig.1. General principle of photoswitchable HaloTag (psHaloTag) and open-closed equilibrium of JF635-HaloTag ligand (HTL). L1 and L2 denote linkers. ]] | ||
The psHaloTag variants show a strong, reversible, deep-red fluorescence turn-on when activated by light. This provides a well-controlled tool for improved imaging. | The psHaloTag variants show a strong, reversible, deep-red fluorescence turn-on when activated by light. This provides a well-controlled tool for improved imaging. | ||
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==== Structural highlights ==== | ==== Structural highlights ==== | ||
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The protein is made by inserting the light-sensing sAsLOV2 domain into the HaloTag protein. An extended helix connects the domains. This connection makes sure the light-induced structural change from the LOV domain is effectively transmitted to the HaloTag region, where the rhodamine dye is bound. Performance-boosting mutations are found near the headgroup of the rhodamine dye. These mutations likely | The protein is made by inserting the light-sensing sAsLOV2 domain into the HaloTag protein. An extended helix connects the domains. This connection makes sure the light-induced structural change from the LOV domain is effectively transmitted to the HaloTag region, where the rhodamine dye is bound. Performance-boosting mutations are found near the headgroup of the rhodamine dye. These mutations likely stabilise the interaction to enhance the fluorescence ON/OFF ratio. | ||
==== Relevance ==== | ==== Relevance ==== | ||
